ZHANG Yi,YU Cun-hao,MA Xiao-hong,et al.Analysis of Isoflavone Synthesis Influenced by UV-B Treatment and Cloning of GmUVR8 Gene in Soybean[J].Soybean Science,2022,41(01):28-035.[doi:10.11861/j.issn.1000-9841.2022.01.0028]
UV-B处理对大豆异黄酮合成影响及GmUVR8基因克隆与表达分析
- Title:
- Analysis of Isoflavone Synthesis Influenced by UV-B Treatment and Cloning of GmUVR8 Gene in Soybean
- Keywords:
- soybean; UV-B; UVR8 photoreceptor; isoflavones; gene expression; gene clone
- 文献标志码:
- A
- 摘要:
- 为了探究UV-B辐射对大豆异黄酮合成调控的分子机理，本研究以UV-B辐射处理大豆V1期幼苗，采用HPLC和定量PCR方法分别测定处理前后各部位异黄酮含量和基因的表达，并克隆出GmUVR8光受体基因。发现UV-B辐射处理大豆V1期幼苗8 h后，叶中总异黄酮含量提高1.3倍，大豆苷元和染料木素分别提高15.8和16.5倍。大豆根、茎和叶中CHS和IFS基因对UV-B反应的时间和强度存在明显差异，CHS11在处理2 h后表达量增加38.4倍，IFS1和IFS2基因的最高表达量分别比处理前增加4.7和18.3倍。克隆出了大豆中编码UVR8光受体的GmUVR8a、GmUVR8b和GmUVR8c基因，其氨基酸序列与拟南芥AtUVR8的同源度为74%；GmUVR8a、GmUVR8b、GmUVR8c基因在叶中表达存在显著差异，以GmUVR8b表达量最高。结果表明在大豆中UV-B辐射可能通过多种UVR8光受体调控苯基丙酸类途径关键酶基因的表达，进而影响异黄酮的合成。
- Abstract:
- In order to explore molecular mechanism of soybean isoflavone synthesis influenced by UV-B radiation， isoflavone accumulation and gene expression levels in different parts of V1 stage, seedlings under UV-B treatment were investigated by HPLC and quantitative real-time PCR respectively， and genes encoding UV RESISTANCE LOCUS 8 (UVR8) photoreceptor were also cloned. The accumulation of total isoflavones in leaves of soybean V1 stage was increased by 1.3 times under 8 h UV-B radiation， and daidzein， genistein improved by 15.8 and 16.5 times respectively. The transcripts of CHS and IFS exhibited obvious difference response to time after UV-B stress， and CHS11 was increased 38.4 times after 2 h treatment， and the highest expression levels of IFS1 and IFS2 increased by 4.7 and 18.3 times respectively. The coding sequences of UVR8 photoreceptor GmUVR8a， GmUVR8b and GmUVR8c were cloned from soybean， the encoding protein had 74% homology in amino acids with AtUVR8， their expression levels in leaves showed significant difference， and GmUVR8b displayed the highest transcripts among those genes. The results indicated that UV-B radiation might regulate synthesis of soybean isoflavones through multiple UVR8 light receptors which affect the expression of key enzymes in phenylpropanoid pathway.
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备注/Memo
收稿日期：2021-06-27